mRNA Amplification Kit

The SMART mRNA Amplification Kit provides a fast and easy method for generating large amounts of sense strand RNA (or mRNA) from limited starting material. If you have inadequate amounts of RNA but need to perform expression, structural, or functional studies, this kit provides a way to amplify your sample while maintaining the relative abundance of transcripts. The SMART mRNA Amplification Kit produces mRNA suitable for quantitative RT-PCR, in vitro translation, cDNA cloning, and array probe generation. SMART mRNA can replace total RNA in any conventional labeling protocol, resulting in lower background, higher signal intensity, and increased dynamic range.

SMART Technology Ensures High-Quality Results

The SMART mRNA amplification method combines reverse transcription with the patented SMART technology to amplify full-length transcripts. First-strand cDNA synthesis is primed by a modified oligo(dT) primer (the CDS Primer II A) that contains additional sequence at the 3’end. The resulting full-length, single-stranded cDNA contains 5' end sequences that are complementary to the SMART T7 oligonucleotide. The SMART T7 anchor sequence is then used for primer extension to generate double-stranded cDNA. Finally, sense strand RNA (mRNA) is transcribed in vitro using T7 RNA polymerase and limiting amounts of ribonucleotides, resulting in linear amplification of mRNA. This method generates large amounts of mRNA for your application in a single round of amplification and preserves the relative gene representation in your sample (1).

Use the Clontech RNA/cDNA Quality Assay to check the integrity of your RNA starting material before beginning your experiment. Because this assay uses RT-PCR, it provides a direct functional test of your sample for its ability to produce full-length cDNA for your application. Achieve quick results using standard lab equipment, and avoid inconvenient and toxic formaldehyde gels.

Applications

References

Chenchik, A., et al. (1998) Generation and use of high-quality cDNA from small amounts of total RNA by SMART PCR. In Gene Cloning and Analysis by RT-PCR (BioTechniques Books, MA), pp. 305–319.

Additional Information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.